半滑舌鳎转化生长因子TGF-β1和TGF-β3基因的克隆及受哈维氏弧菌感染后表达分析  

Molecular Cloning and Expression Analysis of Transforming Growth Factor TGF-β1 and TGF-β3 in Half-smooth Tongue Sole(Cynoglossus semilaevis) Following Stimulation with Vibrio harveyi

作　　者：李雪[1,2,3] 陈松林[1,3] 杨长庚[4] 邵长伟[1,3] 李仰真[1,3] 位战飞 

机构地区：[1]中国水产科学研究院黄海水产研究所/农业部海洋渔业可持续发展重点开放实验室,青岛266071 [2]上海海洋大学水产与生命学院,上海201306 [3]青岛海洋科学与技术国家实验室/海洋渔业科学与食物产出过程功能实验室,青岛266237 [4]中国水产科学研究院长江水产研究所/农业部淡水生物多样性保护与利用重点开放实验室,武汉430223

出　　处：《农业生物技术学报》2016年第6期889-898,共10页Journal of Agricultural Biotechnology

基　　金：国家自然科学基金重点项目(No.31530078);山东省泰山学者攀登计划专项

摘　　要：转化生长因子β(transforming growth factorβ,TGF-β)是一类具有多种功能的蛋白超家族,在细胞免疫、细胞增殖分化和组织损伤的修复中起着关键性作用。本研究从半滑舌鳎(Cynoglossus semilaevis)肝脏中克隆获得了TGF-β1和TGF-β3基因。推导的TGF-β1和TGF-β3氨基酸序列均含有多个N糖基化位点和一个TGF-β家族标签。系统进化树分析显示,半滑舌鳎TGF-β1和TGF-β3分别与鱼类的TGF-β1和TGF-β3亲缘关系最为密切。q RT-PCR结果表明,半滑舌鳎TGF-β1和TGF-β3基因在健康鱼的多个组织中均有表达,二者在皮肤中表达量最高,在肌肉中表达量最低。经哈维氏弧菌(Vibrio harveyi)感染后,TGF-β1在肝脏、脾脏和肾脏中呈现先上升后下降的表达趋势,在感染48 h后的肝脏中表达量达到最大值,是对照组的3.17倍;TGF-β3在脾脏、肾脏和鳃中也呈现先上升后下降的表达趋势,在感染24 h后的鳃中表达量达到最大值,是对照组的4.71倍。以上结果提示,TGF-β1和TGF-β3可能在半滑舌鳎抵御细菌感染的免疫中发挥了重要作用,本研究为证明二者参与机体免疫调节提供了有力证据,为半滑舌鳎分子免疫研究提供了理论依据。Transforming growth factor β （TGF-β） superfamily plays critical roles in immune cell functional regulation, cell proliferation and differentiation, tissue wound repair. The half-smooth tongue sole （Cynoglossus semilaevis）, with excellent properties such as fast growth, individual large, meat delicious and nutrient-rich, is an important economic marine fish species in the North of China. It has become a new kind of excellent breeding object in recent years. But with the enlargement of the farming scale, a variety of infectious diseases is becoming more and more frequently, especially the bacterial disease such as the Vibrio harveyi infection, fast onset and spread quickly. These diseases bring great economic losses to the aquaculture industry. Therefore, in order to carry out the half-smooth tongue sole immune genetic research and explore its immune response mechanism with Vibrio harveyi infection, the TGF-β1 and TGF-β3 genes were cloned from liver in the half -smooth tongue sole. Sequence analysis showed that TGF-β1 and TGF-β3 amino acid sequence consisted of multiple N-glycosylation sites and a TGF-β family signature. Analysis of phylogenetic tree indicated that TGF-β1 and TGF-β3 of the half-smooth tongue sole had a high homology with that of fish. The qRT-PCR analysis showed that TGF-β1 and TGF-β3 expressed in a diverse array of tissues including heart, kidney, intestine, muscle, skin, brain, gill, liver and spleen of healthy half-smooth tongue sole and there were significant differences. The highest levels of TGF-β1 were detected in skin, the higher levels were discovered in kidney and the lowest levels were found in muscle. TGF-β3 expressed the highest in skin, higher in brain and the lowest in muscle. After the Vibrio harveyi infection, the expression levels of TGF-β1 showed uptrend in liver, peaked at 48 h after infection, with 3.17 times of the control group, then fell to control levels. In the spleen, TGF-β1 and TGF-β3 showed similar expression trend , rising in the first stage and

关 键 词：半滑舌鳎 转化生长因子β(TGF-β) TGF-β1 TGF-Β3 表达分析 

分 类 号：S917[农业科学—水产科学]